title: Inhibition of NF-κB activation sensitizes U937 cells to 3'-azido-3'-deoxythymidine induced apoptosis.
abstract: In this study, we investigated molecular mechanisms underlying low susceptibility to apoptosis induced by the nucleoside analog azidothymidine (AZT) and the role of nuclear factor-κB (NF-κB) activation in these phenomena. A preliminary screening in different cell lines indicated U937 monocytic cell line as suitable to this purpose. Treatment of U937 cells even with suprapharmacological concentrations of AZT induced only moderate levels of apoptosis. Surprisingly, SuperArray analysis showed that AZT induced the transcriptional activity of both pro- and anti-apoptotic genes. Interestingly, moreover, several genes upregulated by AZT were NF-κB related. In fact, AZT, after an initial inhibition of NF-κB activation with respect to control, induced a transient, but consistent, increase in NF-κB-binding activity. Inhibition of NF-κB activation in U937 cells, stably transfected with a dominant-negative IκBα or by pharmacological treatment, sensitized them to apoptosis induced by AZT and impaired the upregulation of anti-apoptotic genes in response to AZT treatment, with respect to control cells. These results indicate that NF-κB activation by AZT has a role in protecting target cells from apoptotic cell death, improving our understanding of the toxicology and the therapeutic usage of this drug.
authors.affiliation: Department of Experimental Medicine and Biochemical Sciences, University of Rome 'Tor Vergata', Italy.
authors.forename: C
authors.initials: C
authors.lastname: Matteucci
authors.forename: A
authors.initials: A
authors.lastname: Minutolo
authors.forename: E
authors.initials: E
authors.lastname: Balestrieri
authors.forename: F
authors.initials: F
authors.lastname: Marino-Merlo
authors.forename: P
authors.initials: P
authors.lastname: Bramanti
authors.forename: E
authors.initials: E
authors.lastname: Garaci
authors.forename: B
authors.initials: B
authors.lastname: Macchi
authors.forename: A
authors.initials: A
authors.lastname: Mastino
mesh_terms: D000963:Antimetabolites / Q000494:pharmacology*; D017209:Apoptosis; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D020298:U937 Cells; D015215:Zidovudine / Q000494:pharmacology*
chemical_list: D000963:Antimetabolites; D016328:NF-kappa B; D015215:Zidovudine